Spindle Afferents Research Articles

HCN1 and HCN2 in Rat DRG Neurons: Levels in Nociceptors and Non-Nociceptors, NT3-Dependence and Influence of CFA-Induced Skin Inflammation on HCN2 and NT3 Expression

Ih, which influences neuronal excitability, has recently been measured in vivo in sensory neuron subtypes in dorsal root ganglia (DRGs). However, expression levels of HCN (hyperpolarization-activated cyclic nucleotide-gated) channel proteins that underlie Ih were unknown. We therefore examined immunostaining of the most abundant isoforms in DRGs, HCN1 and HCN2 in these neuron subtypes. This immunostaining was cytoplasmic and membrane-associated (ring). Ring-staining for both isoforms was in neurofilament-rich A-fiber neurons, but not in small neurofilament-poor C-fiber neurons, although some C-neurons showed cytoplasmic HCN2 staining. We recorded intracellularly from DRG neurons in vivo, determined their sensory properties (nociceptive or low-threshold-mechanoreceptive, LTM) and conduction velocities (CVs). We then injected fluorescent dye enabling subsequent immunostaining. For each dye-injected neuron, ring- and cytoplasmic-immunointensities were determined relative to maximum ring-immunointensity. Both HCN1- and HCN2-ring-immunointensities were positively correlated with CV in both nociceptors and LTMs; they were high in Aβ-nociceptors and Aα/β-LTMs. High HCN1 and HCN2 levels in Aα/β-neurons may, via Ih, influence normal non-painful (e.g. touch and proprioceptive) sensations as well as nociception and pain. HCN2-, not HCN1-, ring-intensities were higher in muscle spindle afferents (MSAs) than in all other neurons. The previously reported very high Ih in MSAs may relate to their very high HCN2. In normal C-nociceptors, low HCN1 and HCN2 were consistent with their low/undetectable Ih. In some C-LTMs HCN2-intensities were higher than in C-nociceptors. Together, HCN1 and HCN2 expressions reflect previously reported Ih magnitudes and properties in neuronal subgroups, suggesting these isoforms underlie Ih in DRG neurons. Expression of both isoforms was NT3-dependent in cultured DRG neurons. HCN2-immunostaining in small neurons increased 1 day after cutaneous inflammation (CFA-induced) and recovered by 4 days. This could contribute to acute inflammatory pain. HCN2-immunostaining in large neurons decreased 4 days after CFA, when NT3 was decreased in the DRG. Thus HCN2-expression control differs between large and small neurons.

A dynamical system-Markov model for active postsynaptic responses of muscle spindle afferent nerve

Motoneuron is the control unit of skeletal muscles, and the dynamic frequency-regulating feedback from the afferent nerve of receptors like muscle spindles forms the physical basis of its closed-loop regulation. Focused on the synapses of muscle spindle afferents, this paper established a dynamical system-Markov model starting from presynaptic stimulations to postsynaptic responses, and further verified the model via comparisons between theoretical results and relevant experimental data. With the purpose of describing the active features of dendritic membrane, we employed the methods of dynamical systems rather than the traditional passive cable theory, and identified the physical meaning of parameters involved. For the dynamic behavior of postsynaptic currents, we adopted simplified Markov models so that the analytical solutions for the open dynamics of postsynaptic receptors can be obtained. The model in this paper is capable of simulating the actual non-uniformity of channel density, and is suitable for complex finite element analysis of neurons; thus it facilitates the exploration of the frequency-regulating feedback and control mechanisms of motoneurons.

Tendon vibration attenuates superficial venous vessel response of the resting limb during static arm exercise

BackgroundThe superficial vein of the resting limb constricts sympathetically during exercise. Central command is the one of the neural mechanisms that controls the cardiovascular response to exercise. However, it is not clear whether central command contributes to venous vessel response during exercise. Tendon vibration during static elbow flexion causes primary muscle spindle afferents, such that a lower central command is required to achieve a given force without altering muscle force. The purpose of this study was therefore to investigate whether a reduction in central command during static exercise with tendon vibration influences the superficial venous vessel response in the resting limb.MethodsEleven subjects performed static elbow flexion at 35% of maximal voluntary contraction with (EX + VIB) and without (EX) vibration of the biceps brachii tendon. The heart rate, mean arterial pressure, and rating of perceived exertion (RPE) in overall and exercising muscle were measured. The cross-sectional area (CSAvein) and blood velocity of the basilic vein in the resting upper arm were assessed by ultrasound, and blood flow (BFvein) was calculated using both variables.ResultsMuscle tension during exercise was similar between EX and EX + VIB. However, RPEs at EX + VIB were lower than those at EX (P <0.05). Increases in heart rate and mean arterial pressure during exercise at EX + VIB were also lower than those at EX (P <0.05). CSAvein in the resting limb at EX decreased during exercise from baseline (P <0.05), but CSAvein at EX + VIB did not change during exercise. CSAvein during exercise at EX was smaller than that at EX + VIB (P <0.05). However, BFvein did not change during the protocol under either condition. The decreases in circulatory response and RPEs during EX + VIB, despite identical muscle tension, showed that activation of central command was less during EX + VIB than during EX. Abolishment of the decrease in CSAvein during exercise at EX + VIB may thus have been caused by a lower level of central command at EX + VIB rather than EX.ConclusionDiminished central command induced by tendon vibration may attenuate the superficial venous vessel response of the resting limb during sustained static arm exercise.

Preservation of VGLUT1 synapses on ventral calbindin-immunoreactive interneurons and normal locomotor function in a mouse model of spinal muscular atrophy

Dysfunction in sensorimotor synapses is one of the earliest pathological changes observed in a mouse model [spinal muscular atrophy (SMA)Δ7] of spinal muscular atrophy. Here, we examined the density of proprioceptive and cholinergic synapses on calbindin-immunoreactive interneurons ventral to the lateral motor column. This population includes inhibitory Renshaw interneurons that are known to receive synaptic input from muscle spindle afferents and from motoneurons. At postnatal day (P)13, near the end stage of the disease, the somatic area of calbindin(+) neurons in the L1/L2 and L5/L6 segments was reduced in SMAΔ7 mice compared with controls. In addition, the number and density of terminals expressing the glutamate vesicular transporter (VGLUT1) and the vesicular acetylcholine transporter (VAChT) were increased on calbindin(+) cells in the L1-L2 but not in the L5-L6 segments of SMAΔ7 mice. In addition, the isolated spinal cord of SMA mice was able to generate locomotor-like activity at P4-P6 in the presence of a drug cocktail or in response to dorsal root stimulation. These results argue against a generalized loss of proprioceptive input to spinal circuits in SMA and suggest that the loss of proprioceptive synapses on motoneurons may be secondary to motoneuron pathology. The increased number of VGLUT1(+) and VAChT(+) synapses on calbindin(+) neurons in the L1/L2 segments may be the result of homeostatic mechanisms. Finally, we have shown that abnormal locomotor network function is unlikely to account for the motor deficits observed in SMA mice at P4-6.

Control of position and movement is simplified by combined muscle spindle and Golgi tendon organ feedback

Whereas muscle spindles play a prominent role in current theories of human motor control, Golgi tendon organs (GTO) and their associated tendons are often neglected. This is surprising since there is ample evidence that both tendons and GTOs contribute importantly to neuromusculoskeletal dynamics. Using detailed musculoskeletal models, we provide evidence that simple feedback using muscle spindles alone results in very poor control of joint position and movement since muscle spindles cannot sense changes in tendon length that occur with changes in muscle force. We propose that a combination of spindle and GTO afferents can provide an estimate of muscle-tendon complex length, which can be effectively used for low-level feedback during both postural and movement tasks. The feasibility of the proposed scheme was tested using detailed musculoskeletal models of the human arm. Responses to transient and static perturbations were simulated using a 1-degree-of-freedom (DOF) model of the arm and showed that the combined feedback enabled the system to respond faster, reach steady state faster, and achieve smaller static position errors. Finally, we incorporated the proposed scheme in an optimally controlled 2-DOF model of the arm for fast point-to-point shoulder and elbow movements. Simulations showed that the proposed feedback could be easily incorporated in the optimal control framework without complicating the computation of the optimal control solution, yet greatly enhancing the system's response to perturbations. The theoretical analyses in this study might furthermore provide insight about the strong physiological couplings found between muscle spindle and GTO afferents in the human nervous system.

Jaw muscle spindle afferents coordinate multiple orofacial motoneurons via common premotor neurons in rats: An electrophysiological and anatomical study

Jaw muscle spindle afferents (JMSA) in the mesencephalic trigeminal nucleus (Vme) project to the parvocellular reticular nucleus (PCRt) and dorsomedial spinal trigeminal nucleus (dm-Vsp). A number of premotor neurons that project to the trigeminal motor nucleus (Vmo), facial nucleus (VII) and hypoglossal nucleus (XII) are also located in the PCRt and dm-Vsp. In this study, we examined whether these premotor neurons serve as common relay pool for relaying JMSA to multiple orofacial motoneurons. JMSA inputs to the PCRt and dm-Vsp neurons were verified by recording extracellular responses to electrical stimulation of the caudal Vme or masseter nerve, mechanical stimulation of jaw muscles and jaw opening. After recording, biocytin in recording electrode was inotophorized into recording sites. Biocytin-Iabeled fibers traveled to the Vmo, VII, XII, and the nucleus ambiguus (Amb). Labeled boutons were seen in close apposition with Nissl-stained motoneurons in the Vmo, VII, XII and Amb. In addition, an anterograde tracer (biotinylated dextran amine) was iontophorized into the caudal Vme, and a retrograde tracer (Cholera toxin B subunit) was delivered into either the VII or Xll to identify VII and XII premotor neurons that receive JMSA input. Contacts between labeled Vme neuronal boutons and premotor neurons were observed in the PCRt and adjacent dm-Vsp. Confocal microscopic observations confirmed close contacts between Vme boutons and VII and XII premotor neurons. This study provides evidence that JMSA may coordinate activities of multiple orofacial motor nuclei, including Vmo, VII, XII and Amb in the brainstem via a common premotor neuron pool.

Expression and properties of hyperpolarization-activated current in rat dorsal root ganglion neurons with known sensory function.

The hyperpolarization-activated current (Ih) has been implicated in nociception/pain, but its expression levels in nociceptors remained unknown. We recorded Ih magnitude and properties by voltage clamp from dorsal root ganglion (DRG) neurons in vivo, after classifying them as nociceptive or low-threshold-mechanoreceptors (LTMs) and as having C-, Aδ- or Aα/β-conduction velocities (CVs). For both nociceptors and LTMs, Ih amplitude and Ih density (at −100 mV) were significantly positively correlated with CV. Median Ih magnitudes and Ih density in neuronal subgroups were respectively: muscle spindle afferents (MSAs): −4.6 nA, −33 pA pF−1; cutaneous Aα/β LTMs: −2.2 nA, −20 pA pF−1; Aβ-nociceptors: −2.6 nA, −21 pA pF−1; both Aδ-LTMs and nociceptors: −1.3 nA, ∼−14 pA pF−1; C-LTMs: −0.4 nA, −7.6 pA pF−1; and C-nociceptors: −0.26 nA, −5 pA pF−1. Ih activation slow time constants (slow τ values) were strongly correlated with fast τ values; both were shortest in MSAs. Most neurons had τ values consistent with HCN1-related Ih; others had τ values closer to HCN1+HCN2 channels, or HCN2 in the presence of cAMP. In contrast, median half-activation voltages (V0.5) of −80 to −86 mV for neuronal subgroups suggest contributions of HCN2 to Ih. τ values were unrelated to CV but were inversely correlated with Ih and Ih density for all non-MSA LTMs, and for Aδ-nociceptors. From activation curves ∼2–7% of Ih would be activated at normal membrane potentials. The high Ih may be important for excitability of A-nociceptors (responsible for sharp/pricking-type pain) and Aα/β-LTMs (tactile sensations and proprioception). Underlying HCN expression in these subgroups therefore needs to be determined. Altered Ih expression and/or properties (e.g. in chronic/pathological pain states) may influence both nociceptor and LTM excitability.

Requirement of neuronal connexin36 in pathways mediating presynaptic inhibition of primary afferents in functionally mature mouse spinal cord

Electrical synapses formed by gap junctions containing connexin36 (Cx36) promote synchronous activity of interneurones in many regions of mammalian brain; however, there is limited information on the role of electrical synapses in spinal neuronal networks. Here we show that Cx36 is widely distributed in the spinal cord and is involved in mechanisms that govern presynaptic inhibition of primary afferent terminals. Electrophysiological recordings were made in spinal cord preparations from 8- to 11-day-old wild-type and Cx36 knockout mice. Several features associated with presynaptic inhibition evoked by conditioning stimulation of low threshold hindlimb afferents were substantially compromised in Cx36 knockout mice. Dorsal root potentials (DRPs) evoked by low intensity stimulation of sensory afferents were reduced in amplitude by 79% and in duration by 67% in Cx36 knockouts. DRPs were similarly affected in wild-types by bath application of gap junction blockers. Consistent with presynaptic inhibition of group Ia muscle spindle afferent terminals on motoneurones described in adult cats, conditioning stimulation of an adjacent dorsal root evoked a long duration inhibition of monosynaptic reflexes recorded from the ventral root in wild-type mice, and this inhibition was antagonized by bicuculline. The same conditioning stimulation failed to inhibit monosynaptic reflexes in Cx36 knockout mice. Immunofluorescence labelling for Cx36 was found throughout the dorsal and ventral horns of the spinal cord of juvenile mice and persisted in mature animals. In deep dorsal horn laminae, where interneurones involved in presynaptic inhibition of large diameter muscle afferents are located, cells were extensively dye-coupled following intracellular neurobiotin injection. Coupled cells displayed Cx36-positive puncta along their processes. Our results indicate that gap junctions formed by Cx36 in spinal cord are required for maintenance of presynaptic inhibition, including the regulation of transmission from Ia muscle spindle afferents. In addition to a role in presynaptic inhibition in juvenile animals, the persistence of Cx36 expression among spinal neuronal populations in the adult mouse suggests that the contribution of electrical synapses to integrative processes in fully mature spinal cord may be as diverse as that found in other areas of the CNS.

Paw-shake response and locomotion: can one CPG generate two different rhythmic behaviors?

Rhythmic limb movements like locomotion or paw-shake response are controlled by network of spinal circuits, known as central pattern generators (CPGs), as evidenced from locomotor-like and paw-shake like activity in limb peripheral nerves elicited in decerebrate or spinal animals with blocked neuromuscular transmission [4]. Unlike fictive locomotion and scratch, that are likely controlled by distinct CPGs [3], fictive paw-shake response has not been systematically investigated and it is not known whether it is controlled by a specialized CPG or by the CPG that also controls locomotion. In-vivo recordings of paw-shake motor patterns elicited by stimulation of paw skin afferents [7] have revealed high frequency hindlimb oscillations (~10 Hz) with atypical muscle synergies – reciprocal activation of anterior and posterior hindlimb muscles in each half of the paw-shake cycle; both anterior and posterior muscle groups include flexor and extensor muscles. We asked whether a paw-shake response with the atypical muscle synergies can be generated by a typical half-center locomotor CPG reciprocally activating flexor and extensor muscles. Using software AnimatLab [2] we developed a 5-segment cat hindlimb model with 12 Hill-type muscle actuators controlled by (1) a half-center CPG activating flexor and extensor muscles (two-joint muscles received both flexion- and extension-related signals [5,6]) and (2) proprioceptive input originated from the muscle spindle and Golgi tendon organ afferents. The CPG was modeled by two single-compartment spiking neurons in a half-center configuration. Other neurons (Ia-afferents, alpha-motor neurons, Ia-interneurons, and interneurons mediating autogenic and heterogenic reflex pathways) were modeled as non-spiking neurons (firing rate model based on work by [1]). Model parameters were adjusted such that computer simulations reproduced the recorded paw-shake mechanics and the anterior-posterior muscle activation patterns. The obtained results demonstrated that a half-center locomotor CPG can produce movement mechanics and muscle activity patterns typical for paw-shake responses if (1) the locomotor CPG is capable to operate at frequencies 3 to10 times higher than during locomotion and (2) synaptic weights in spinal circuits can be modified during paw-shake response. We speculate that the two conditions can be realized by sensory input from paw skin afferents.

The posterior insular-opercular region and the search of a primary cortex for pain

To be considered specific for nociception, a cortical region should: (a) have plausible connections with ascending nociceptive pathways; (b) be activated by noxious stimuli; (c) trigger nociceptive sensations if directly stimulated; and (d) tone down nociception when injured. In addition, lesions in this area should have a potential to develop neuropathic pain, as is the case of all lesions in nociceptive pathways. The single cortical region approaching these requirements in humans encompasses the suprasylvian posterior insula and its adjoining medial operculum (referred to as "PIMO" in this review). This region does not contain, however, solely nociceptive networks, but represents in primates the main sensory receiving area of the spinothalamic system, and as such contributes to the processing of thermo-sensory, nociceptive, C-fibre tactile, and visceral input. Nociception (and, a fortiori, pain) should therefore not be considered as a separate sensory modality, like vision or audition, but rather as one component of a global system subtending the most primitive forms of somatosensation. Although a clear functional segregation of PIMO sub-areas has not yet been achieved, some preferential distribution has been described in humans: pain-related networks appear preferentially distributed within the posterior insula, and non-noxious thermal processing in the adjacent medial operculum. Thus, spinothalamic sub-modalities may be partially segregated in the PIMO, in analogy with the separate representation of dorsal column input from joint, muscle spindle and tactile afferents in S1. Specificity, however, may not wholly depend on ascending 'labelled lines' but also on cortical network properties driven by intrinsic and extrinsic circuitry. Given its particular anatomo-functional properties, thalamic connections, and tight relations with limbic and multisensory cortices, the PIMO region deserves to be considered as a third somatosensory region (S3) devoted to the processing of spinothalamic inputs.

Characterization of Muscle Spindle Afferents in the Adult Mouse Using an In Vitro Muscle-Nerve Preparation

We utilized an in vitro adult mouse extensor digitorum longus (EDL) nerve-attached preparation to characterize the responses of muscle spindle afferents to ramp-and-hold stretch and sinusoidal vibratory stimuli. Responses were measured at both room (24°C) and muscle body temperature (34°C). Muscle spindle afferent static firing frequencies increased linearly in response to increasing stretch lengths to accurately encode the magnitude of muscle stretch (tested at 2.5%, 5% and 7.5% of resting length [Lo]). Peak firing frequency increased with ramp speeds (20% Lo/sec, 40% Lo/sec, and 60% Lo/sec). As a population, muscle spindle afferents could entrain 1:1 to sinusoidal vibrations throughout the frequency (10–100 Hz) and amplitude ranges tested (5–100 µm). Most units preferentially entrained to vibration frequencies close to their baseline steady-state firing frequencies. Cooling the muscle to 24°C decreased baseline firing frequency and units correspondingly entrained to slower frequency vibrations. The ramp component of stretch generated dynamic firing responses. These responses and related measures of dynamic sensitivity were not able to categorize units as primary (group Ia) or secondary (group II) even when tested with more extreme length changes (10% Lo). We conclude that the population of spindle afferents combines to encode stretch in a smoothly graded manner over the physiological range of lengths and speeds tested. Overall, spindle afferent response properties were comparable to those seen in other species, supporting subsequent use of the mouse genetic model system for studies on spindle function and dysfunction in an isolated muscle-nerve preparation.

Discharge rates and discharge variability of muscle spindle afferents in human chronic spinal cord injury

ObjectivesTo test the hypothesis that the firing rates and discharge variability of human muscle spindles are not affected by spinal cord injury. MethodsTungsten microelectrodes were inserted into muscle fascicles of the peroneal nerve in six individuals with complete paralysis of the lower limbs following spinal cord injury: 12 afferents were spontaneously active at rest and 7 were recruited during passive muscle stretch. For comparison, recordings were made from 17 spontaneously active and 9 stretch-recruited afferents in 12 intact subjects. ResultsFiring rates for the spontaneously active muscle spindles were not significantly different between the spinal (9.8±1.6Hz) and intact (10.2±1.3Hz) subjects; the same was true for the stretch-recruited afferents – static firing rates, measured over the final 1s of a ramp-and-hold stretch, were not different between the spinal and intact groups (13.1±3.1% vs 10.0±2.5Hz). There were also no differences in discharge variability between the spinal and intact subjects, either for the spontaneously active spindles (8.1±2.0% vs 5.7±0.9%) or for the stretch-activated spindles, calculated over the final 1s of static stretch (19.7±5.6% vs 17.0±1.9%). In addition, the responses to stretch imposed manually by the experimenter provided no evidence for an increase in the dynamic response to stretch in the patients. ConclusionsThe static stretch sensitivity of human muscle spindles is not affected by chronic spinal cord injury, suggesting that there is no difference in static (and possibly dynamic) fusimotor drive to paralyzed muscles in chronic spinal cord injury. SignificanceThis study provides no evidence for an increase in fusimotor drive as a mechanism for the spasticity associated with chronic spinal injury, though further studies using controlled stretch would be required before it can be concluded that dynamic fusimotor drive is “normal” in these patients.

The decreased responsiveness of lumbar muscle spindles to a prior history of spinal muscle lengthening is graded with the magnitude of change in vertebral position

In the lumbar spine, muscle spindle responsiveness is affected by the duration and direction of a lumbar vertebra’s positional history. The purpose of the present study was to determine the relationship between changes in the magnitude of a lumbar vertebra’s positional history and the responsiveness of lumbar muscle spindles to a subsequent vertebral position and subsequent vertebral movement. Neural activity from multifidus and longissimus muscle spindle afferents in deeply anesthetized cats was recorded while creating positional histories of the L6 vertebra. History was induced using a displacement-controlled feedback motor. It held the L6 vertebra for 4s at an intermediate position (hold-intermediate at 0mm) and at seven positions from 0.07 to 1.55mm more ventralward and dorsalward which lengthened (hold-long) and shortened (hold-short) the lumbar muscles. Following the conditioning hold positions, L6 was returned to the intermediate position. Muscle spindle discharge at this position and during a lengthening movement was compared between hold-intermediate and hold-short conditionings and between hold-intermediate and hold-short conditionings. We found that regardless of conditioning magnitude, the seven shortening magnitudes similarly increased muscle spindle responsiveness to both vertebral position and movement. In contrast, the seven lengthening magnitudes produced a graded decrease in responsiveness to both position and movement. The decrease to position became maximal following conditioning magnitudes of ∼0.75mm. The decrease to movement did not reach a maximum even with conditioning magnitudes of ∼1.55mm. The data suggest that the fidelity of proprioceptive information from muscle spindles in the low back is influenced by small changes in the previous length history of lumbar muscles.

Muscle spindle responses to horizontal support surface perturbation in the anesthetized cat: insights into the role of autogenic feedback in whole body postural control

Intact cats and humans respond to support surface perturbations with broadly tuned, directionally sensitive muscle activation. These muscle responses are further sensitive to initial stance widths (distance between feet) and perturbation velocity. The sensory origins driving these responses are not known, and conflicting hypotheses are prevalent in the literature. We hypothesize that the direction-, stance-width-, and velocity-sensitive muscle response during support surface perturbations is driven largely by rapid autogenic proprioceptive pathways. The primary objective of this study was to obtain direct evidence for our hypothesis by establishing that muscle spindle receptors in the intact limb can provide appropriate information to drive the muscle response to whole body postural perturbations. Our second objective was to determine if spindle recordings from the intact limb generate the heightened sensitivity to small perturbations that has been reported in isolated muscle experiments. Maintenance of this heightened sensitivity would indicate that muscle spindles are highly proficient at detecting even small disturbances, suggesting they can provide efficient feedback about changing postural conditions. We performed intraaxonal recordings from muscle spindles in anesthetized cats during horizontal, hindlimb perturbations. We indeed found that muscle spindle afferents in the intact limb generate broadly tuned but directionally sensitive activation patterns. These afferents were also sensitive to initial stance widths and perturbation velocities. Finally, we found that afferents in the intact limb have heightened sensitivity to small perturbations. We conclude that muscle spindle afferents provide an array of important information about biomechanics and perturbation characteristics highlighting their potential importance in generating appropriate muscular response during a postural disturbance.

P01.25. Classification of L6 muscle spindle afferents in the anesthesized cat

Purpose Patients with low back pain demonstrate proprioceptive difficulties including spinal repositioning errors and impaired lumbosacral proprioceptive acuity. Few data are available regarding proprioceptive properties of muscle spindles in the low back. Muscle spindle afferents can originate from 2 types of receptive endings which terminate and respond to mechanical changes in 3 types of intrafusal fibers. Receptive endings may be primary or secondary. They terminate on dynamic bag (b1) static bag (b2 )a nd/or chain (c) intrafusal fibers. We sought to classify lumbar paraspinal muscle spindle afferents based upon their receptive endings and intrafusal terminations. Classification was based on their responses to ramp and hold vertebral movement before and after intrafusal activation using succinylcholine (SCH, 100-300ug/kg.ia). Afferents terminating in primary endings are especially responsive to the dynamic ramp stimulus. During intrafusal activation, afferents terminating on b1 fibers further increase their discharge to the dynamic ramp whereas those terminating on b2 fibers increase their static resting discharge. Methods Electrophysiological recordings from spindle afferents (n=195) were obtained from L6 dorsal root filaments with receptive fields in the L6 longissimus and multifidus muscles in an anesthetized cat preparation. Controlled vertebral actuations that stretched the paraspinal muscles were applied to the L6 spinous process in a dorsal-ventralward direction [1.5mm (n=120), 1.6mm (n=21), or 1.7mm (n=54) using a feedback motor system. Instantaneous discharge frequency was averaged and compared over three ramp cycles pre- and postSCH injection.

Frequency-dependent amplification of stretch-evoked excitatory input in spinal motoneurons.

Voltage-dependent calcium and sodium channels mediating persistent inward currents (PICs) amplify the effects of synaptic inputs on the membrane potential and firing rate of motoneurons. CaPIC channels are thought to be relatively slow, whereas the NaPIC channels have fast kinetics. These different characteristics influence how synaptic inputs with different frequency content are amplified; the slow kinetics of Ca channels suggest that they can only contribute to amplification of low frequency inputs (<5 Hz). To characterize frequency-dependent amplification of excitatory postsynaptic potentials (EPSPs), we measured the averaged stretch-evoked EPSPs in cat medial gastrocnemius motoneurons in decerebrate cats at different subthreshold levels of membrane potential. EPSPs were produced by muscle spindle afferents activated by stretching the homonymous and synergist muscles at frequencies of 5-50 Hz. We adjusted the stretch amplitudes at different frequencies to produce approximately the same peak-to-peak EPSP amplitude and quantified the amount of amplification by expressing the EPSP integral at different levels of depolarization as a percentage of that measured with the membrane hyperpolarized. Amplification was observed at all stretch frequencies but generally decreased with increasing stretch frequency. However, in many cells the amount of amplification was greater at 10 Hz than at 5 Hz. Fast amplification was generally reduced or absent when the lidocaine derivative QX-314 was included in the electrode solution, supporting a strong contribution from Na channels. These results suggest that NaPICs can combine with CaPICs to enhance motoneuron responses to modulations of synaptic drive over a physiologically significant range of frequencies.

Modulation of Human Muscle Spindle Discharge by Arterial Pulsations - Functional Effects and Consequences

Arterial pulsations are known to modulate muscle spindle firing; however, the physiological significance of such synchronised modulation has not been investigated. Unitary recordings were made from 75 human muscle spindle afferents innervating the pretibial muscles. The modulation of muscle spindle discharge by arterial pulsations was evaluated by R-wave triggered averaging and power spectral analysis. We describe various effects arterial pulsations may have on muscle spindle afferent discharge. Afferents could be “driven” by arterial pulsations, e.g., showing no other spontaneous activity than spikes generated with cardiac rhythmicity. Among afferents showing ongoing discharge that was not primarily related to cardiac rhythmicity we illustrate several mechanisms by which individual spikes may become phase-locked. However, in the majority of afferents the discharge rate was modulated by the pulse wave without spikes being phase locked. Then we assessed whether these influences changed in two physiological conditions in which a sustained increase in muscle sympathetic nerve activity was observed without activation of fusimotor neurones: a maximal inspiratory breath-hold, which causes a fall in systolic pressure, and acute muscle pain, which causes an increase in systolic pressure. The majority of primary muscle spindle afferents displayed pulse-wave modulation, but neither apnoea nor pain had any significant effect on the strength of this modulation, suggesting that the physiological noise injected by the arterial pulsations is robust and relatively insensitive to fluctuations in blood pressure. Within the afferent population there was a similar number of muscle spindles that were inhibited and that were excited by the arterial pulse wave, indicating that after signal integration at the population level, arterial pulsations of opposite polarity would cancel each other out. We speculate that with close-to-threshold stimuli the arterial pulsations may serve as an endogenous noise source that may synchronise the sporadic discharge within the afferent population and thus facilitate the detection of weak stimuli.

Acute inflammation alters the response to stretch in mouse extensor digitorum longus muscle afferents

Inflammatory pain alters muscle spindle responses to stretch (Wenngren, et al, 1998; Ro &amp; Capra, 2001), but the exact mechanism of sensitization is unknown. We used an in vitro mouse hindlimb muscle‐nerve preparation to study plasticity in afferent responses to ramp and hold stretches following acute inflammation (0.5 mg/kg lipopolysaccharide (LPS) 18 hrs before experiment). Following LPS, units had lower baseline firing and more units were only active during stretch (6/16 LPS vs. 3/26 control). The LPS units also displayed a significantly greater decrease in firing frequency over the 4 s stretch. Some LPS units (3/16) are likely group III/IV instead of muscle spindle afferents; they had very small responses to stretch and did not show the characteristic frequency adaptation. Overall, the acute systemic inflammatory state induced by LPS clearly alters the response profile of muscle afferents to stretch, and therefore proprioceptive signaling. Further study on these changes are planned to provide insight into underlying mechanisms, their behavioral relevance, and to identify strategies that normalize function.Supported by K12 GM000680 NIH/NIGMS More Division (KAW) and Pfizer (SH).

Origin of the low-level EMG during the silent period following transcranial magnetic stimulation

ObjectiveThe cortical silent period refers to a period of near silence in the electromyogram (EMG) after transcranial magnetic stimulation (TMS) of the motor cortex during contraction. However, low-level EMG of unknown origin is often present. We hypothesised that it arises through spinal reflexes. Sudden lengthening of the muscle as force drops during the silent period could excite muscle spindles and facilitate motoneurones. MethodsSubjects (n=8) performed maximal isometric, shortening and lengthening contractions of the elbow flexors during which TMS (90–100% output) was delivered over the motor cortex. The rate of flexion during shortening contractions reduced muscle lengthening caused by muscle relaxation. Surface EMG was recorded from biceps brachii and brachioradialis, and the low-level EMG during silent periods produced by TMS was measured. ResultsLow-level EMG activity was reduced on average by 68% in biceps and 63% in brachioradialis in the shortening contraction compared to all other contraction conditions (p<0.001). Levels of pre-stimulus EMG were similar between conditions. ConclusionsMuscle lengthening contributes to low-level EMG activity in the silent period, through spinal reflex facilitation by muscle spindle afferents. SignificanceThe silent period depth is not only dependent on cortical output but also reflex effects evoked by muscle lengthening.

Can loss of muscle spindle afferents explain the ataxic gait in Riley-Day syndrome?

The Riley-Day syndrome is the most common of the hereditary sensory and autonomic neuropathies (Type III). Among the well-recognized clinical features are reduced pain and temperature sensation, absent deep tendon reflexes and a progressively ataxic gait. To explain the latter we tested the hypothesis that muscle spindles, or their afferents, are absent in hereditary sensory and autonomic neuropathy III by attempting to record from muscle spindle afferents from a nerve supplying the leg in 10 patients. For comparison we also recorded muscle spindles from 15 healthy subjects and from two patients with hereditary sensory and autonomic neuropathy IV, who have profound sensory disturbances but no ataxia. Tungsten microelectrodes were inserted percutaneously into fascicles of the common peroneal nerve at the fibular head. Intraneural stimulation within muscle fascicles evoked twitches at normal stimulus currents (10-30 µA), and deep pain (which often referred) at high intensities (1 mA). Microneurographic recordings from muscle fascicles revealed a complete absence of spontaneously active muscle spindles in patients with hereditary sensory and autonomic neuropathy III; moreover, responses to passive muscle stretch could not be observed. Conversely, muscle spindles appeared normal in patients with hereditary sensory and autonomic neuropathy IV, with mean firing rates of spontaneously active endings being similar to those recorded from healthy controls. Intraneural stimulation within cutaneous fascicles evoked paraesthesiae in the fascicular innervation territory at normal stimulus intensities, but cutaneous pain was never reported during high-intensity stimulation in any of the patients. Microneurographic recordings from cutaneous fascicles revealed the presence of normal large-diameter cutaneous mechanoreceptors in hereditary sensory and autonomic neuropathy III. Our results suggest that the complete absence of functional muscle spindles in these patients explains their loss of deep tendon reflexes. Moreover, we suggest that their ataxic gait is sensory in origin, due to the loss of functional muscle spindles and hence a compromised sensorimotor control of locomotion.